Effects of cordycepin on airway inflammatory cell recruitment in response to intratracheal administration of LPS in mice. C57BL/6 mice (n ≥ 5) were subjected to a single i.p. injection of 2 mg/kg cordycepin (suspended in ethanol and diluted 1/10 in saline) or vehicle 1 h before the intratracheal administration of LPS (50 μg/mouse) suspended in 50 μL saline. Either 6 or 24 h after treatment, mice were euthanized and subjected to either lung collection for RNA analysis or BAL, respectively. (A) BAL fluids were collected and centrifuged; cells were then differentially stained and visualized by light microscopy followed by a count of total cells and neutrophils. Data are given as the means ± SEM of values obtained from at least five mice per group. ∗Different from untreated control mice; P < 0.01. #Different from LPS-treated mice; P< 0.01. (B) Whole lungs were collected and subjected to RNA extraction followed by cDNA generation. Prepared cDNA was subjected to real-time PCR with primers for VCAM-1, ICAM-1, MCP-1, MIP-1α, MIP-2, KC or CXCR2; β-actin was amplified by its specific primers and used as an internal control and on which the attained values were normalized (C) The cDNA from the different experimental groups described above was assessed for TNF-α by real-time PCR. ∗Different from untreated control mice; P< 0.01; #Different from LPS-treated mice; P < 0.01.